反相高效液相色谱测定庆大霉素效价方法的研究

Determination of Gentamicin Titres by RP-HPLC

建立了反相高效液相色谱(Reverse Phase liquid Chromatography,RP-HPLC)测定庆大霉素效价的方法,结合管碟法所做的生物效价进行比较,其相关度R^2=0.9823,RSD=1.29,证明RP-HPLC可直接测定庆大霉素的效价,从而解决了生测效价程序复杂,人为误差大的问题。RP-HPLC测定方法:采用岛津高效液相色谱SPD-10AVP、UV-VIS、LC-10ATVP,PepMapC_(18) ^(Tm)4.6×150 mmSilica C18(5μm300 A)Phenomenex ODS色谱柱,流动相MILLI-Q水:冰醋酸:甲醇(比例为25:5:70)配置0.02mol/L庚烷磺酸钠溶液,流速1.0 mL/min,检测波长330 nm,检测灵敏度0.020 AUFS,柱温为室温,进样量20μL,在10 min内可对样品液中C_1、C_(1a)、C_(2a)、C_2 4种物质同时进行定性定量测定。

In the paper a new method was established by RP-HPLC for determination of gentamicin titres. The new method compared with the biological assay the correlation R2 = 0. 9823, RSD = 1.29. So it could be determinated gentamicin titres direietely and the problem by bioassay that process was complicate, error was severely was solved. RP-HPLC method：high-performance liquid chromatography Shimadzu SPD- 10AVP,UV-VIS,LC-10ATVP,PepMapC18Tm4. 6 x 150 mm Silica C18 （5 μm 300 A） Phenomenex ODS column, mobile phase was prepared with O. 02 mol / L of heptane sodium by the MILLI-Q water：glacial acetic acid：methanol （25： 5： 70） ,flow rate 1.0 mL / min,detection at 330 nm, detection sensitivity 0. 020AUFS, temperature of column was room temperature, injection volume 20 μL. The quality and quantity of C1 ,Cla ,C2a ,C2 in the sample fluid were measured at the same time for 10 min.